PMID- 10438503
OWN - NLM
STAT- MEDLINE
DCOM- 19990901
LR  - 20190508
IS  - 0021-9258 (Print)
IS  - 0021-9258 (Linking)
VI  - 274
IP  - 33
DP  - 1999 Aug 13
TI  - Human acyl-CoA:cholesterol acyltransferase-1 in the endoplasmic reticulum
      contains seven transmembrane domains.
PG  - 23276-85
AB  - Acyl-CoA:cholesterol acyltransferase (ACAT) plays important roles in cellular
      cholesterol homeostasis and is involved in atherosclerosis. ACAT-1 protein is
      located mainly in the ER. The hydropathy plot suggests that ACAT-1 protein
      contains multiple transmembrane segments. We inserted either the hemagglutinin
      tag or the HisT7 tag at various hydrophilic regions within the human ACAT-1
      protein and used immunofluorescence microscopy to determine the topography of the
      tagged proteins expressed in mutant Chinese hamster ovary cells lacking
      endogenous ACAT. All of the tagged proteins are located mainly in the ER and
      retain full or partial enzyme activities. None of the tagged proteins produces
      detectable intracellular degradation intermediates. Treating cells with digitonin
      at 5 micrograms/ml permeabilizes the plasma membranes while leaving the ER
      membranes sealed; in contrast, treating cells with 0.25% Triton X-100 or with
      cold methanol permeabilizes both the plasma membranes and the ER membranes. After
      appropriate permeabilization, double immunostaining using antibodies against the 
      N-terminal region and against the inserted tag were used to visualize various
      regions of the tagged protein. The results show that human ACAT-1 in the ER
      contains seven transmembrane domains.
FAU - Lin, S
AU  - Lin S
AD  - Department of Biochemistry, Dartmouth Medical School, Hanover, New Hampshire
      03755, USA.
FAU - Cheng, D
AU  - Cheng D
FAU - Liu, M S
AU  - Liu MS
FAU - Chen, J
AU  - Chen J
FAU - Chang, T Y
AU  - Chang TY
LA  - eng
GR  - HL 36709/HL/NHLBI NIH HHS/United States
GR  - HL 60306/HL/NHLBI NIH HHS/United States
PT  - Journal Article
PT  - Research Support, U.S. Gov't, P.H.S.
PL  - United States
TA  - J Biol Chem
JT  - The Journal of biological chemistry
JID - 2985121R
RN  - 0 (DNA Primers)
RN  - 0 (Membrane Proteins)
RN  - 0 (Recombinant Proteins)
RN  - EC 2.3.1.26 (Sterol O-Acyltransferase)
SB  - IM
MH  - Algorithms
MH  - Amino Acid Sequence
MH  - Animals
MH  - Base Sequence
MH  - CHO Cells
MH  - Cricetinae
MH  - DNA Primers
MH  - Endoplasmic Reticulum/*enzymology
MH  - Humans
MH  - Membrane Proteins/chemistry/*metabolism
MH  - Microscopy, Fluorescence
MH  - Molecular Sequence Data
MH  - Recombinant Proteins/metabolism
MH  - Sterol O-Acyltransferase/chemistry/*metabolism
EDAT- 1999/08/07 00:00
MHDA- 1999/08/07 00:01
CRDT- 1999/08/07 00:00
PHST- 1999/08/07 00:00 [pubmed]
PHST- 1999/08/07 00:01 [medline]
PHST- 1999/08/07 00:00 [entrez]
AID - 10.1074/jbc.274.33.23276 [doi]
PST - ppublish
SO  - J Biol Chem. 1999 Aug 13;274(33):23276-85. doi: 10.1074/jbc.274.33.23276.